Most outbreaks of Shiga toxin–producing Escherichia coli (STEC) are attributed to consumption of contaminated foodstuffs including beef and dairy products. In this study, we evaluated the prevalence of non-O157 STEC cultured from beef and dairy cattle and collected in Xinjiang Uygur Autonomous Region in China. Results identified 67 non-O157 STEC recovered from the 793 samples including beef cattle (10.28%, 43/418) and dairy cattle (6.40%, 24/375). A total of 67 non-O157 STEC was sequenced allowing for in silico analyses of their serotypes, virulence genes, and identification of the corresponding multilocus sequence types (STs). Twenty-one O serogroups and nine H serotypes were identified and the dominant serotype identified was O22:H8. One stx1 subtype (stx1a) and four stx2 subtypes (2a, 2b, 2c, and 2d) were found in the 67 non-O157 STEC isolates. The results revealed that stx1a+stx2a-positive STEC isolates were predominant (32.83%, 22/67), followed by stx1a+stx2d (29.85%, 20/67) and stx2a alone (17.91%, 12/67). Non-O157 STEC isolates carried virulence genes ehxA (98.51%), subA (53.73%), and cdtB (17.91%). Of the four adherence-associated genes tested, eaeA was absent, whereas lpfA and iha were present in 67 and 55 non-O157 STEC isolates, respectively. The STEC isolates were divided into 48 pulsed-field gel electrophoresis patterns and 10 STs, and ST446 (O22:H8) was the dominant clone (22.38%). Our results revealed that there was a high genetic diversity among non-O157 STEC isolated from beef and dairy cattle, some of which have potential to cause human diseases.
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References
1.
AdamuMS, UgochukwuICI, IdokoSI, KwabuggeYA, AbubakarNS, AmehJA. Virulent gene profile and antibiotic susceptibility pattern of Shiga toxin-producing Escherichia coli (STEC) from cattle and camels in Maiduguri, North-Eastern Nigeria. Trop Anim Health Prod, 2018; 50:1327–1341.
2.
AfsetJE, BruantG, BrousseauR, HarelJ, AnderssenE, BevangerL, BerghK. Identification of virulence genes linked with diarrhea due to atypical enteropathogenic Escherichia coli by DNA microarray analysis and PCR. J Clin Microbiol, 2006; 44:3703–3711.
3.
AlonsoCA, MoraA, DíazD, BlancoM, González-BarrioD, Ruiz-FonsF, SimónC, BlancoJ, TorresC. Occurrence and characterization of stx and/or eae-positive Escherichia coli isolated from wildlife, including a typical EPEC strain from a wild boar. Vet Microbiol, 2017; 207:69–73.
4.
BaiL, GuoY, LanR, DongY, WangW, HuY, GanX, YanS, FuP, PeiXJFC. Genotypic characterization of Shiga toxin-producing Escherichia coli O157:H7 isolates in food products from china between 2005 and 2010. Food Control, 2015; 50:209–214.
5.
BaiX, HuB, XuY, SunH, ZhaoA, BaP, FuS, FanR, JinY, WangH, GuoQ, XuX, LuS, XiongY. Molecular and phylogenetic characterization of non-O157 Shiga toxin-producing Escherichia coli strains in China. Front Cell Infect Microbiol, 2016; 6:143.
6.
BankevichA, NurkS, AntipovD, GurevichAA, DvorkinM, KulikovAS, LesinVM, NikolenkoSI, PhamS, PrjibelskiAD, PyshkinAV, SirotkinAV, VyahhiN, TeslerG, AlekseyevMA, PevznerPA. SPAdes: A new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol, 2012; 19:455–477.
7.
BeutinL, StrauchE. Identification of sequence diversity in the Escherichia coli fliC genes encoding flagellar types H8 and H40 and its use in typing of Shiga toxin-producing E. coli O8, O22, O111, O174, and O179 strains. J Clin Microbiol, 2007; 45:333–339.
8.
BielaszewskaM, MellmannA, ZhangW, KöckR, FruthA, BauwensA, PetersG, KarchH. Characterisation of the Escherichia coli strain associated with an outbreak of haemolytic uraemic syndrome in Germany, 2011: A microbiological study. Lancet Infect Dis, 2011; 11:671–676.
9.
BoerlinP, McEwenSA, Boerlin-PetzoldF, WilsonJB, JohnsonRP, GylesCL. Associations between virulence factors of Shiga toxin-producing Escherichia coli and disease in humans. J Clin Microbiol, 1999; 37:497–503.
CabalA, PorreroMC, de la CruzML, SaezJL, BarcenaC, LopezG, GortazarC, DominguezL, AlvarezJ. Molecular characterization and antimicrobial resistance of STEC strains isolated from healthy cattle in 2011 and 2013 in Spain. Epidemiol Infect, 2016; 144:2956–2966.
12.
Castanie-cornetMP, PenfoundTA, SmithD, ElliottJF, FosterJW. Control of acid resistance in Escherichia coli. J Bacteriol, 1999; 181:3525–3535.
13.
ChartH, SmithHR, ScotlandSM, RoweB, MilfordDV, TaylorCM. Serological identification of Escherichia coli O157:H7 infection in haemolytic uraemic syndrome. Lancet, 1991; 337:138–140.
14.
DingH, SuZQ, XiaLN, WangYM, WangD, ZhangJY, ZhangT, KuangL, YaoG. Influence of immunomagnetic enrichment method for isolation and identification of E. coli O157:H7 from bovine. China Anim Husband Vet Med, 2017; 44:1189–1194.
15.
EFSA European Food Safety Authority. Scientific report on the European Union One Health 2018 Zoonoses Report. EFSA J, 2019; 17:276.
16.
EFSA. Scientific Opinion on the pathogenicity assessment of Shiga toxin-producing Escherichia coli (STEC) and the public health risk posed by contamination of food with STEC. EFSA J, 2020; 18:5105.
FAO and WHO. Attributing Illness Caused by ShigaToxin-Producing. Escherichia coli. (STEC) to Specific Foods. Microbiological Risk Assessment Series No. 32, Rome. p. 74. ISSN: 1726-5274.2019.
19.
FAO and WHOSTEC EXPERT GROUP. Hazard identification and characterization: Criteria for categorizing Shiga toxin-producing Escherichia coli on a risk basis (dagger). J Food Prot, 2019; 82:7–21.
20.
FengPCH, ReddyS. Prevalences of Shiga toxin subtypes and selected other virulence factors among Shiga-toxigenic Escherichia coli strains isolated from fresh produce. Appl Environ Microbiol, 2013; 79:6917–6923.
21.
FerdousM, FriedrichAW, GrundmannH, de BoerRF, CroughsPD, IslamMA, Kluytmans-van den BerghMF, Kooistra-SmidAM, RossenJW. Molecular characterization and phylogeny of Shiga toxin-producing Escherichia coli isolates obtained from two Dutch regions using whole genome sequencing. Clin Microbiol Infect, 2016; 22:642.e641–e649.
22.
FranzolinMR, AlvesRC, KellerR, GomesTA, BeutinL, BarretoML, MilroyC, StrinaA, RibeiroH, TrabulsiLR. Prevalence of diarrheagenic Escherichia coli in children with diarrhea in Salvador, Bahia, Brazil. Mem Inst Oswaldo Cruz, 2005; 100:359–363.
23.
FratamicoPM, YanX, LiuY, DebRoyC, ByrneB, MonaghanA, FanningS, BoltonD. Escherichia coli serogroup O2 and O28ac O-antigen gene cluster sequences and detection of pathogenic E. coli O2 and O28ac by PCR. Can J Microbiol, 2010; 56:308–316.
24.
FuSS, BaiXN, FanRY, XuYM, XuXB, XiongYW. Molecular characteristics of human-derived non-O157 Shiga toxin-producing Escherichia coli strains isolated in five regions of China. Chin J Microbiol Immunol, 2017; 37:213–218.
25.
GalarceN, EscobarB, SánchezF, Paredes-OssesE, Alegría-MoránR, BorieC, BorieC. Virulence genes, Shiga toxin subtypes, serogroups, and clonal relationship of Shiga toxin-producing Escherichia coli strains isolated from livestock and companion animals. Animals (Basel), 2019; 9:733.
26.
González-EscalonaN, KaseJA. Virulence gene profiles and phylogeny of Shiga toxin-positive Escherichia coli strains isolated from FDA regulated foods during 2010–2017. PLoS One, 2019; 14:e0214620.
27.
HadlerJL, ClogherP, HuangJ, LibbyT, CronquistA, WilsonS, RyanP, SaupeA, NicholsonC, McGuireS, ShiferawB, DunnJ, HurdS. The relationship between census tract poverty and Shiga toxin-producing E. coli risk, analysis of FoodNet Data, 2010–2014. Open Forum Infect Dis, 2018; 5:ofy148.
28.
HassanJ, NazirKHMNH, ParvejMS, KamalT, RahmanMT. Molecular based prevalence of shiga toxigenic Escherichia coli in rectal swab of apparently healthy cattle in Mymensingh district, Bangladesh. J Adv Vet Anim Res, 2017; 4:194–199.
29.
HauserE, MellmannA, SemmlerT, StoeberH, WielerLH, KarchH, KueblerN, FruthA, HarmsenD, WenigerT, TietzeE, SchmidtH. Phylogenetic and molecular analysis of food-borne shiga toxin-producing Escherichia coli. Appl Environ Microbiol, 2013; 79:2731–2740.
30.
HusseinHS. Prevalence and pathogenicity of Shiga toxin-producing Escherichia coli in beef cattle and their products. J Anim Sci, 2007; 85:E63–E72.
31.
HusseinHS, BollingerLM. Prevalence of Shiga toxin-producing Escherichia coli in beef cattle. J Food Prot, 2005; 68:2224–2241.
32.
HusseinHS, SakumaT. Prevalence of Shiga toxin-producing Escherichia coli in dairy cattle and their products. J Dairy Sci, 2005; 88:450–465.
33.
InouyeM, DashnowH, RavenLA, SchultzMB, PopeBJ, TomitaT, ZobelJ, HoltKE. SRST2: Rapid genomic surveillance for public health and hospital microbiology labs. Genome Med, 2014; 6:90.
34.
IsikoJ, KhaitsaM, BergholzTM. Novel sequence types of non-O157 Shiga toxin-producing Escherichia coli isolated from cattle. Lett Appl Microbiol, 2015; 60:552–557.
35.
JajarmiM, Imani FooladiAA, BadoueiMA, AhmadiA. Virulence genes, Shiga toxin subtypes, major O-serogroups, and phylogenetic background of Shiga toxin-producing Escherichia coli strains isolated from cattle in Iran. Microb Pathog, 2017; 109:274–279.
36.
JinnerotT, TomaselliATP, JohannessenGS, SöderlundR, UrdahlAM, AspánA, SekseC. The prevalence and genomic context of Shiga toxin 2a genes in E. coli found in cattle. PLoS One, 2020; 15:e0232305.
37.
JoensenKG, ScheutzF, LundO, HasmanH, KaasRS, NielsenEM, AarestrupFM. Real-time whole-genome sequencing for routine typing, surveillance, and outbreak detection of verotoxigenic Escherichia coli. J Clin Microbiol, 2014; 52:1501–1510.
38.
JoensenKG, TetzschnerAM, IguchiA, AarestrupFM, ScheutzF. Rapid and easy in silico serotyping of Escherichia coli isolates by sse of whole-genome sequencing data. J Clin Microbiol, 2015; 53:2410–2426.
39.
JostM, MäusezahlM, BloombergG, StephanR. Serotypes and virulence profiles of Shiga toxin-producing Escherichia coli strains isolated during 2017 from human infections in Switzerland. Int J Med Microbiol, 2018; 308:933–939.
40.
KangE, HwangSY, KwonKH, KimKY, KimJH, ParkYH. Prevalence and characteristics of Shiga toxin-producing Escherichia coli (STEC) from cattle in Korea between 2010 and 2011. J Vet Sci, 2014; 15:369–379.
KimJ-S, LeeM-S, KimJH. Recent updates on outbreaks of Shiga toxin-producing Escherichia coli and its potential reservoirs. Front Cell Infect Microbiol, 2020; 10:273.
43.
LiY, LuoQ, ShiX, LinY, QiuY, LvD, JiangY, ChenQ, JiangM, MaH, ChengJ, HuQ. Phenotypic and genotypic characterization of clinical enterotoxigenic Escherichia coli isolates from Shenzhen, China. Foodborne Pathog Dis, 2017; 14:333–340.
44.
MaingaAO, Cenci-GogaBT, MalahlelaMN, TshumaT, KalakeA, KaramaM. Occurrence and characterization of seven major Shiga toxin-producing Escherichia coli serotypes from healthy cattle on cow-calf operations in South Africa. Zoonoses Public Health, 2018; 65:777–789.
45.
MajowiczSE, ScallanE, Jones-BittonA, SargeantJM, StapletonJ, AnguloFJ, YeungDH, KirkMD. Global incidence of human Shiga toxin–producing Escherichia coli infections and deaths: A systematic review and knowledge synthesis. Foodborne Pathog Dis, 2014; 11:447–455.
46.
MarçaisG, DelcherAL, PhillippyAM, CostonR, SalzbergSL, ZiminA. MUMmer4: a fast and versatile genome alignment system. PLoS Comput Biol, 2018; 14:e1005944.
47.
MartorelliL, AlbaneseA, VilteD, CantetR, BentancorA, ZolezziG, ChinenI, IbarraC, RivasM, MercadoEC, CataldiA. Shiga toxin-producing Escherichia coli (STEC) O22:H8 isolated from cattle reduces E. coli O157:H7 adherence in vitro and in vivo. Vet Microbiol, 2017; 208:8–17.
48.
MekataH, IguchiA, KawanoK, KirinoY, KobayashiI, MisawaN. Identification of O serotypes, genotypes, and virulotypes of Shiga toxin–producing Escherichia coli isolates, including non-O157 from beef cattle in Japan. J Food Prot, 2014; 77:1269–1274.
MonteroDA, CantoFD, VelascoJ, ColelloR, PadolaNL, SalazarJC, MartinCS, OnateA, BlancoJ, RaskoDA, ContrerasC, PuenteJL, ScheutzF, FranzE, VidalRM. Cumulative acquisition of pathogenicity islands has shaped virulence potential and contributed to the emergence of LEE-negative Shiga toxin-producing Escherichia coli strains. Emerg Microbes Infect, 2019; 8:486–502.
52.
MurindaSE, NguyenLT, LandersTL, DraughonFA, MathewAG, HoganJS, SmithKL, HancockDD, OliverSP. Comparison of Escherichia coli Isolates from humans, food, and farm and companion animals for presence of Shiga toxin-producing E. coli virulence markers. Foodborne Pathog Dis, 2004; 1:178–184.
53.
MyliusM, DreesmanJ, PulzM, PallaschG, BeyrerK, ClaußenK, AllerbergerF, FruthA, LangC, PragerR, FliegerA, SchlagerS, KalhöferD, MertensE. Shiga toxin-producing Escherichia coli O103:H2 outbreak in Germany after school trip to Austria due to raw cow milk, 2017—The important role of international collaboration for outbreak investigations. Int J Med Microbiol, 2018; 308:539–544.
54.
OrthD, GrifK, KhanAB, NaimA, DierichMP, WürznerR. The Shiga toxin genotype rather than the amount of Shiga toxin or the cytotoxicity of Shiga toxin in vitro correlates with the appearance of the hemolytic uremic syndrome. Diagn Microbiol Infect Dis, 2007; 59:235–242.
55.
PatonAW, SrimanoteP, WoodrowMC, PatonJC. Characterization of Saa, a novel autoagglutinating adhesin produced by locus of enterocyte effacement-negative Shiga-toxigenic Escherichia coli strains that are virulent for humans. Infect Immun, 2001; 69:6999–7009.
56.
PengZ, LiangW, HuZ, LiX, GuoR, HuaL, TangX, TanC, ChenH, WangX, WuB. O-serogroups, virulence genes, antimicrobial susceptibility, and MLST genotypes of Shiga toxin-producing Escherichia coli from swine and cattle in Central China. BMC Vet Res, 2019; 15:427.
57.
PriceMN, DehalPS, ArkinAP. FastTree: Computing large minimum evolution trees with profiles instead of a distance matrix. Mol Biol Evol, 2009; 26:1641–1650.
58.
PugsleyAP, SchwartzM. Expression of a gene in a 400-base-pair fragment of colicin plasmid ColE2-P9 is sufficient to cause host cell lysis. J Bacteriol, 1983; 156:109–114.
59.
RibotEM, FairMA, GautomR, CameronDN, HunterSB, SwaminathanB, BarrettTJ. Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne Pathog Dis, 2006; 3:59–67.
60.
RileyLW, RemisRS, HelgersonSD, McGeeHB, WellsJG, DavisBR, HebertRJ, OlcottES, JohnsonLM, HargrettNT, BlakePA, CohenML. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med, 1983; 308:681–685.
61.
RossBN, MaricarmenRL, CiezaRJ, McwilliamsBD, TorresAG. The role of long polar fimbriae in Escherichia coli O104:H4 adhesion and colonization. PLoS One, 2015; 10:e0141845.
62.
ScallanE, HoekstraRM, AnguloFJ, TauxeRV, WiddowsonMA, RoySL, JonesJL, GriffinPM. Foodborne illness acquired in the United States—Major pathogens. Emerg Infect Dis, 2011; 17:7–15.
63.
ScheutzF. Taxonomy meets public health: The case of Shiga toxin-producing Escherichia coli. Microbiol Spectr, 2014; 2. DOI: 10.1128/microbiolspec.EHEC-0019-2013.
64.
ScheutzF, TeelLD, BeutinL, PiérardD, BuvensG, KarchH, MellmannA, CaprioliA, TozzoliR, MorabitoS, StrockbineNA, Melton-CelsaAR, SanchezM, PerssonS, O'BrienAD. Multicenter evaluation of a sequence-based protocol for subtyping Shiga toxins and standardizing Stx nomenclature. J Clin Microbiol, 2012; 50:2951–2963.
65.
ShridharPB, PatelIR, GangiredlaJ, NollLW, ShiXR, BaiJF, ElkinsCA, StrockbineNA, NagarajaTG. Genetic analysis of virulence potential of Escherichia coli O104 serotypes isolated from cattle feces using whole genome sequencing. Front Microbiol, 2018; 9:341.
SoysalN, Mariani-KurkdjianP, SmailY, LiguoriS, GoualiM, LoukiadisE, FachP, BruyandM, BlancoJ, BidetP, BonacorsiS. Enterohemorrhagic Escherichia coli hybrid pathotype O80:H2 as a new therapeutichallenge. Emerg Infect Dis, 2016; 22:1604–1612.
68.
SuZQ, WangD, ZhangJY, ZhangY, LiuYY, MaKQ, GaoJJ, SunX, YaoG. Study on Seasonal Impact to the Distribution of Bovine Escherichia coli O157∶H7 in Xinjiang. China Anim Husband Vet Med, 2017; 45:212–217.
69.
TackDM, MarderEP, GriffinPM, CieslakPR, DunnJ, HurdS, ScallanE, LathropS, MuseA, RyanP, SmithK, Tobin-D'AngeloM, VugiaDJ, HoltKG, WolpertBJ, TauxeR, GeisslerAL. Preliminary incidence and trends of infections with pathogens transmitted commonly through Food-Foodborne Diseases Active Surveillance Network, 10 U.S. Sites, 2015–2018. MMWR Morb Mortal Wkly Rep, 2019; 68:369–373.
TristãoLCS, GonzalezAGM, CoutinhoCAS, CerqueiraAMF, GomesMJP, IrinoK, GuthBEC, AndradeJRC. Virulence markers and genetic relationships of Shiga toxin-producing Escherichia coli strains from serogroup O111 isolated from cattle. Vet Microbiol, 2007; 119:358–365.
72.
WerberD, ScheutzF. The importance of integrating genetic strain information for managing cases of Shiga toxin-producing E. coli infection. Epidemiol Infect, 2019; 147:e264.
73.
XiongYW, WangP, LanRT, YeCY, WangH, RenJ, JingHQ, WangYT, ZhouZM, Bai XM; CuiZG, LuoX, ZhaoAL, WangY, ZhangSM, SunH, WangL, XuJG. A novel Escherichia coli O157:H7 clone causing a major hemolytic uremic syndrome outbreak in China. PLoS One, 2012; 7:e36144.
74.
YamazakiM, SaitoM, InuzukaK, ShimaS, TaniwakiH, ItoK. eaeA genes in Escherichia coli derived from Japanese patients with sporadic diarrhea. Kansenshogaku Zasshi, 1997; 71:1059–1065.
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